LOCUS       sequence-270186-        6671 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prey vector AT5G59680 O7_pECIA14 should be used with bait vector 
            AT5G59680 O7_pECIA2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6671)
  AUTHORS   Smakowska-Luzan E, Mott GA, Parys K, Stegmann M, Howton TC, 
            Layeghifard M, Neuhold J, Lehner A, Kong J, Grunwald K, Weinberger 
            N, Satbhai SB, Mayer D, Busch W, Madalinski M, Stolt-Bergner P, 
            Provart NJ, Mukhtar MS, Zipfel C, Desveaux D, Guttman DS, Belkhadir 
            Y
  TITLE     An extracellular network of Arabidopsis leucine-rich repeat receptor
            kinases.
  JOURNAL   Nature. 2018 Jan 18;553(7688):342-346. doi: 10.1038/nature25184. 
            Epub 2018 Jan 10.
  PUBMED    29320478
REFERENCE   2  (bases 1 to 6671)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/nature25184"; journalName: "Nature"; date: "2018-01-18- 
            18"; volume: "553"; issue: "7688"; pages: "342-346"
COMMENT     Sequence Label: AT5G59680 O7_pECIA14
FEATURES             Location/Qualifiers
     source          1..6671
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        23..449
                     /label=MT promoter
                     /note="Drosophila metallothionein promoter"
     sig_peptide     467..520
                     /codon_start=1
                     /product="Drosophila BiP signal sequence"
                     /label=BiP signal sequence
                     /translation="MKLCILLAVVAFVGLSLG"
     CDS             1967..1990
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3665..3688
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3704..3721
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3744..3761)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3794..3928
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3818..3837)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3872..3891
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4072..4088)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4072..4088)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4085..4107)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4096..4112
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4120..4126,4127..4144,4145..4150))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4165..4186
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4303..4320)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4474..5062)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4554..4573)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5233..6024,6025..6093))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5856..5875
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6094..6198)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6266..6284
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6322..6344)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6444..6463
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(6657..6671,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 gtaaaacgac ggccagtgaa ttaattcgtt gcaggacagg atgtggtgcc cgatgtgact
       61 agctctttgc tgcaggccgt cctatcctct ggttccgata agagacccag aactccggcc
      121 ccccaccgcc caccgccacc cccatacata tgtggtacgc aagtaagagt gcctgcgcat
      181 gccccatgtg ccccaccaag agctttgcat cccatacaag tccccaaagt ggagaaccga
      241 accaattctt cgcgggcaga acaaaagctt ctgcacacgt ctccactcga atttggagcc
      301 ggccggcgtg tgcaaaagag gtgaatcgaa cgaaagaccc gtgtgtaaag ccgcgtttcc
      361 aaaatgtata aaaccgagag catctggcca atgtgcatca gttgtggtca gcagcaaaat
      421 caagtgaatc atctcagtgc aactaaaggg gggatccgat ctcaatatga agttatgcat
      481 attactggcc gtcgtggcct ttgttggcct ctcgctcggg agccaacaag ggttcatcag
      541 tttggattgt ggtctacccg cgaatgaacc atctccttat acagagccga ggactggatt
      601 gcagttttct tcggacgcag cattcatcca gagtggaaaa attggtagga ttcaagcaaa
      661 tctggaggct gacttcctga agccatcaac gacgatgaga tattttccag acggaaaacg
      721 gaactgctac aatctaaacg ttgagaaggg aagaaatcat cttatcaggg ctcggtttgt
      781 atatggaaat tatgatggtc gtgacactgg ccccaagttt gatctgtatc ttggacctaa
      841 tccatgggcc acgatagatt tggcaaaaca agtgaatggt acacgtcccg agatcatgca
      901 catcccaaca tcaaacaagt tgcaggtttg tcttgttaag actggggaaa ctacaccgct
      961 aatatcggtt ttggaagtac ggccgatggg aagtggtact tatctcacca agtctggctc
     1021 tctgaaactt tactatcgag aatattttag caagtctgat tcttctctaa ggtaccctga
     1081 tgatatctat gaccgccaat ggacttcgtt ctttgataca gagtggacac aaattaacac
     1141 cactagcgat gtgggaaatt ccagtgacta taagccacca aaagtcgctc tcactactgc
     1201 ggccattcca actaatgcca gtgcgccatt gacgaatgaa tggagttcag taaaccccga
     1261 tgaacagtat tacgtatacg cccatttttc tgagatccaa gagttgcagg caaatgaaac
     1321 cagggaattc aacatgttac taaatgggaa actttttttt ggccctgtgg ttcctccaaa
     1381 gctagcaata agtactatcc taagtgtgtc gccgaatact tgtgaaggag gggagtgcaa
     1441 tttacagctg ataagaacca acagatcaac tcttccacct ctacttaacg cttatgaggt
     1501 ctacaaagtt attcagtttc cgcagttgga aacaaatgaa accgacgttt ctgctgtgaa
     1561 aaacatccaa gccacctatg aattgagtag aatcaactgg caaagcgatc catgcgtccc
     1621 tcaacagttt atgtgggatg gtttaaactg cagcatcaca gatataacca cgccaccaag
     1681 aatcactact ttaaacttgt catcaagtgg tttaactgga acgataacag ctgccattca
     1741 aaatcttacg acactagaaa agctggactt gtcgaataat aacttgaccg gagaggtgcc
     1801 agaatttcta agcaacatga aatcgctgtt ggtcataaac ttaagcggga acgatcttaa
     1861 cggtacaatt cctcaatctc tacaaaggaa aggacttgag ctactttatc aaggaaaccc
     1921 caggcttatt tcccctggtt caactgaaac caaatctggc aagagcctgg aggtgctgtt
     1981 ccagggacca agcggcaatc ccggcggcga tctggccccc cagatgctgc gcgagctgca
     2041 ggagaccaat gccgccctgc aggacgtgcg cgagctgctg cgccaccgcg tgaaggagat
     2101 caccttcctg aagaacaccg tgatggagtg cgacgcctgc ggcatgcagc cctctagagg
     2161 ctccggctcc atcatcccag ttgaggagga gaacccggac ttctggaacc gcgaggcagc
     2221 cgaggccctg ggtgccgcca agaagctgca gcctgcacag acagccgcca agaacctcat
     2281 catcttcctg ggcgatggga tgggggtgtc tacggtgaca gctgccagga tcctaaaagg
     2341 gcagaagaag gacaaactgg ggcctgagat acccctggcc atggaccgct tcccatatgt
     2401 ggctctgtcc aagacataca atgtagacaa acatgtgcca gacagtggag ccacagccac
     2461 ggcctacctg tgcggggtca agggcaactt ccagaccatt ggcttgagtg cagccgcccg
     2521 ctttaaccag tgcaacacga cacgcggcaa cgaggtcatc tccgtgatga atcgggccaa
     2581 gaaagcaggg aagtcagtgg gagtggtaac caccacacga gtgcagcacg cctcgccagc
     2641 cggcacctac gcccacacgg tgaaccgcaa ctggtactcg gacgccgacg tgcctgcctc
     2701 ggcccgccag gaggggtgcc aggacatcgc tacgcagctc atctccaaca tggacattga
     2761 cgtgatccta ggtggaggcc gaaagtacat gtttcgcatg ggaaccccag accctgagta
     2821 cccagatgac tacagccaag gtgggaccag gctggacggg aagaatctgg tgcaggaatg
     2881 gctggcgaag cgccagggtg cccggtatgt gtggaaccgc actgagctca tgcaggcttc
     2941 cctggacccg tctgtgaccc atctcatggg tctctttgag cctggagaca tgaaatacga
     3001 gatccaccga gactccacac tggacccctc cctgatggag atgacagagg ctgccctgcg
     3061 cctgctgagc aggaaccccc gcggcttctt cctcttcgtg gagggtggtc gcatcgacca
     3121 tggtcatcat gaaagcaggg cttaccgggc actgactgag acgatcatgt tcgacgacgc
     3181 cattgagagg gcgggccagc tcaccagcga ggaggacacg ctgagcctcg tcactgccga
     3241 ccactcccac gtcttctcct tcggaggcta ccccctgcga gggagctcca tcttcgggct
     3301 ggcccctggc aaggcccggg acaggaaggc ctacacggtc ctcctatacg gaaacggtcc
     3361 aggctatgtg ctcaaggacg gcgcccggcc ggatgttacc gagagcgaga gcgggagccc
     3421 cgagtatcgg cagcagtcag cagtgcccct ggacgaagag acccacgcag gcgaggacgt
     3481 ggcggtgttc gcgcgcggcc cgcaggcgca cctggttcac ggcgtgcagg agcagacctt
     3541 catagcgcac gtcatggcct tcgccgcctg cctggagccc tacaccgcct gcgacctggc
     3601 gccccccgcc ggcaccaccg acgccgcgca cccgggttct agaggaagca cctccggcga
     3661 taacgattac aaggatgatg atgataaggc cagcggcacc ggtcatcatc accatcacca
     3721 ttgagtttaa acccgctgat cagcctcgac tgtgccttct aaggcctgag ctcgctgatc
     3781 agcctcgatc gaggatccag acatgataag atacattgat gagtttggac aaaccacaac
     3841 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt
     3901 aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca
     3961 ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtat
     4021 ggctgattat gatcagtcga cctgcaggca tgcaagcttg gcgtaatcat ggtcatagct
     4081 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     4141 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     4201 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     4261 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     4321 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     4381 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     4441 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     4501 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     4561 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     4621 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     4681 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     4741 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     4801 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     4861 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     4921 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     4981 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     5041 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     5101 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     5161 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     5221 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     5281 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     5341 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     5401 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     5461 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     5521 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     5581 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     5641 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     5701 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     5761 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     5821 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     5881 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     5941 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     6001 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     6061 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     6121 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     6181 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
     6241 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg
     6301 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
     6361 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
     6421 gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat
     6481 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc
     6541 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
     6601 gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca
     6661 gtcacgacgt t
//
